GSM7658555: DAY15_CHIR; Homo sapiens; RNA-Seq
hPSC-derived cardiac cells at day15 were dissociated to single cells by 4 mg/ml collagenase type 4 treatment in PBS for 20 minutes at 37°C and then Accutase treatment for 15 minutes at 37°C. Dissociated cells were stained with DAPI and then live cells were sorted using MoFlo Astrios EQS. Libraries were performed according to the manufacturer's instructions (Single Cell 3' Reagent Kit v3 protocol, 10x Genomics). Briefly, cardiac cells were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Illumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the appropriately sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Illumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.